Translationally controlled tumor protein (TCTP) is an important regulator involved in plant growth and development as well as responses to biotic and abiotic stresses. By taking use of Puccinia triticina isolate-260 and four wheat near-isogenic lines (NILs) that form combinations with different affinity degrees, we investigated the expression of TCTP at transcription and translational levels using RT-qPCR and Western Blotting, respectively. The transcriptional level of TCTP was significantly up-regulated in the incompatible combination, but this was diverse in NILs with Lr gene. No obvious change trend in the affinity combination was observed. Gained from the results of TCTP mRNA and protein levels, the expression of TCTP was found to be modified at both transcription and translation level during the interaction between wheat and Puccinia triticina. It is speculated that it may play a positive role in the defense response of wheat against Puccinia triticina infection. This will lay the foundation for further research on the function and mechanism of TCTP.